Tissue Culture Literation
| Plant latin name | Rehmannia glutiosa Libosch. f. hueichingensis (Chao et Schih) Hsiao |
| Literature code | Rehmannia_glutinosa-Rep-1 |
| Reference | Park SU et al., Journal of Medicinal Plants Research, 3(1): 031-034 (2009) |
| Summary | The establishment of an efficient protocol for plant regeneration and micropropagation from leaf cultures of Rehmannia glutinosa L. is reported. The regenerated shoots obtained from leaf cultures on solid MS medium containing different concentractions of TDZ. The highest number of shoots per explant (2.1) and shoot growth (1.2 cm) was obtained on MS medium containing 1 mg/L TDZ. The addition of 0.1 mg/L NAA and 3g/L Gelrite in MS medium containing 1 mg/L TDZ substantially improved the shoot regeneration of R. glutinosa. The rooted plants were hardened and transferred to soil with a 73% survival rate. The continuous production of R. glutinosa regenerated plants could be used as a possible micropropagation system. |
| Objectives | Development of a method for shoot organogenesis and plant regeneration from leaf explant cultures of R. glutinosa |
| Materials | |
| Explant | Young leaves of R. glutinosa taken from greenhouse grown plants |
| Initial culture | |
| Shoot multiplication | Leaf explants (7 X 7 mm2 in size) were prepared from surface-sterilized leaf, were cultured on MS medium supplenented with different concentrations of TDZ. For improvement of shoot regeneration, the medium was optimized by testing the effect of different concentration of auxins and gelling agents on shoot formation and growth. Cultures were maintained at 25±1 ªC in a growth chamber with a 16-h photoperiod under standard cool white fluorescent tubes (35 µmol s-1 m-2). MS medium containing 1 mg/l TDZ, 0.1 mg/l NAA and 3 g/l Gelrite gave the highest number of shoots per explant (4.7) with highest shoot length (1.8 cm). |
| Rooting | Regenerated shoots (around 1 cm long) were placed in MS medium. The medium was solidified with 3 g/L Gelrite and dispensed at 30 ml per Magenta box and four shoots were cultured in each box. Regenerated shoots were incubated at 25±1ªC in a growth chamber with a 16-h photoperiod under standard cool white fluorescent tubes (35 mmol s-1 m-2) for 8 weeks for rooting of regenerated shoots. |
| Acclimation | The rooted plants were washed with sterile water to remove Gelrite, transferred to pots containing autoclaved vermiculite, and coveredwith polyethylene bags for one week to maintain high humidity. |
| Planting | The acclimataized plants were then transferred to soil and maintained in a growth chamber with a 16-h photoperiod, and a night/day temperature of 18/20ªC for 2 weeks. These hardened plants then transferred to the greenhouse. |
| Cultivation conditions | |
| Traints of regenerants | The rooted plants were hardened and transferred to soil with a 73% survival rate where they grew normally and flowered within 3 months. The continuous production of R. glutinosa regenerated plants could be used as a possible micropropagation system. Every year the growers have to save some roots from total harvested roots of their plant for conventional method for mass propagation, but we can produce about 5 plantlets from in vitro one leaf explant cultures of R. glutinosa all the year round. |
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